EP0163393A2 - Procédé de mise en oeuvre d'un essai immunologique d'agglutination - Google Patents
Procédé de mise en oeuvre d'un essai immunologique d'agglutination Download PDFInfo
- Publication number
- EP0163393A2 EP0163393A2 EP85302453A EP85302453A EP0163393A2 EP 0163393 A2 EP0163393 A2 EP 0163393A2 EP 85302453 A EP85302453 A EP 85302453A EP 85302453 A EP85302453 A EP 85302453A EP 0163393 A2 EP0163393 A2 EP 0163393A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- agglutination
- acid
- additive
- particles
- serum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/815—Test for named compound or class of compounds
- Y10S436/817—Steroids or hormones
- Y10S436/818—Human chorionic gonadotropin
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/825—Pretreatment for removal of interfering factors from sample
Definitions
- the present invention is concerned with assay procedures and more particularly withagglutination immunoassays for an analyte in a fluid sample.
- agglutination immunoassays continue to be widely used in biology and medicine for the detection of small quantities of an antibody or antigen of interest in a fluid test sample.
- the agglutination reaction involves the in vitro aggregation of microscopic carrier particles. This aggregation is mediated by the specific reaction between antibodies and antigens, one of which is immobilized on the surface of the carrier particles.
- a fluid containing the ligand of interest is introduced into a suspension of the sensitized carrier particles and the appearance of aggregation is noted as indicative of the ligand.
- agglutination assays One especially valuable use of the agglutination assays is in the detection of a ligand or analyte of interest in human body fluids such as serum.
- the agglutination reaction may then be used in several different modes to detect an antigen or antibody (the ligand of interest) as follows:
- This reaction mixture is then combined with the antigen immobilized carrier particles.
- the degree to which the ligand of interest (the antigen) in the test sample inhibits the aggregation of the carrier particles that would otherwise have occurred, indicates the concentration of ligand present in the sample;
- a fixed quantity of antigen is mixed with a dilution of the test sample containing the ligand of interest - a specific antibody - which inactivates a portion of the antigen. This reaction mixture is then combined with the antibody immobilized carrier particles.
- latex immunoassay methods for determining the presence of a ligand of interest in a non-extracted body fluid by including a chemical additive comprised of at least one halogen substituted carboxylic acid or a salt of the acid in the reaction mixture to decrease non-specific interferences of the latex immunoassay.
- the additive reduces non-specific interferences in individual serum samples, pooled serum samples and other body fluids for analysis including plasma, saliva, spinal fluid, urine and the like.
- the addition of such additive permits the use of agglutination immunoassays to determine quantitative levels of endogenous metabolites and to monitor specific ligands of interest such as drugs, therapeutic agents and specific binding proteins in the samples.
- the present invention provides a method of performing an agglutination immunoassay for an analyte in a fluid sample which is combined with a reagent which includes sensitized latex particles and a binding partner as required, whereby the degree of agglutination of the particle is a measure of the analyte in the sample, characterised in that non-specific interferences of the agglutination of the particles are decreased by adding to the reaction mixture an additive comprising at least one halogen substituted carboxylic acid or water soluble salt thereof wherein the acid has the formula (I): and wherein R 1 is Cl, Br or I and R . is H, Cl, Br or I and R 3 is H, CH 3 -, CH 3 CH 2 -, CI, Br or I.
- the addition of the compound of Formula I, e.g., sodium trichloroacetate (hereinafter NaTCA) in agglutination reaction mixtures substantially reduces or eliminates completely the effects of non-specific inhibitors and other interferences in serum and non-serum bodily fluid test samples.
- the preferred reagents are chloroacetic acid, dichloroacetic acid, trichloroacetic acid, bromoacetic acid, dibromoacetic acid, tribromoacetic acid, iodoacetic acid or mixtures thereof with NaTCA being the most preferred additive.
- the additive is suitably used in a latex agglutination immunoassay (hereinafter LIA) in which a suspension of latex particles ranging from 0.05-1.0 micrometers in diameter covalently bind or tightly absorb a specific binding partner (an antigen or antibody) for the ligand to be determined in the serum sample.
- LIA latex agglutination immunoassay
- the chemical additive preferably NaTCA is included in the reaction mixture; the addition of the chemical additive will result in an increased level of specificity in the agglutination at the time of measurement.
- the increased degree of agglutination may then be determined visually or be measured using conventional procedures such as turbidimetry, nephelometry, conventional light scattering .techniques, quasielastic scattering methods or angular anisotropic scattering determination.
- NaTCA used as an additive to agglutination mixtures substantially reduces or eliminates all the effects of non-specific serum interferences.
- the use of such salts of halogen substituted carboxylic acids as chemical additives in immunoassay methods allows the user to monitor ligands of interest in serum or plasma samples including drugs, antibiotics and other therapeutic agents.
- a metallic salt of trifluoroacetic acid is not suitable as an additive to reduce non-specific interferences.
- the chemical additive used in this invention can, in fact, be a mixture of two or more metallic salts of halogen substituted carboxylic acids.
- a mixture of sodium or potassium trichloroacetate and sodium (or potassium) tribromoacetate is effective as an additive. It is preferred that the pH of the additive be maintained between 8.0 and 10.0 for maximum effect. This range may be adjusted to accommodate any optimum immunochemical proportions in the individual test system.
- the optimum concentration of the additive depends on numerous factors which include a) latex particle composition, b) nature of the antibody and its sensitivity to the additive, c) nature of the antigen or napten and its sensitivity to the additive and d) temperature - all of which define the system.
- the optimum concentration of the additive must be determined empirically for each system. Conceivably there are some systems which may be completely incompatible with the additive for reasons having nothing to do with interference by serum.
- Agglutination mixtures (0.4 ml, pH 8.0) which contained 0.02 M sodium barbital, 0.15 M NaCl, 0.1% bovine serum albumin, a 1/92 dilution of latex particles coated with hCG (Pregnosis® latex available from Roche Diagnostics, Nutley, NJ), and serially diluted anti-human chorionic gonadotropin (anti-hCG) antiserum (Miles-Yeda, No 67-073) were prepared. Mixtures which included, in addition, 0.44 M NaTCA or 5.0% normal male serum (hCG- free) or both were also prepared.
- the agglutination mixtures were maintained at 25°C and turbidimetric measurements performed in a spectophoto- meter. Readings of the absorbance at 600 nm were made immediately after combining all components and again after a 30 minute incubation. The change in absorbance during that interval for each mixture was calculated by subtracting the zero time reading from the-30 minute reading. Additionally, appropriate blanks in which antibody was absent were run in parallel, and any normal change in absorbance was subtracted, resulting in a ⁇ A value for each mixture (Table I).
- Incubation mixtures 350 ⁇ l, pH 8.0 which contained 1/7,000 dilution of anti-hCG antiserum (1/3,500 dilution in mixtures containing NaTCA), 0.02 M sodium barbital, 0.15 M NaCl, 0.1% bovine serum albumin and varying amounts of hCG were prepared. Other mixtures which included, in addition, 0.53 M TCA, 5.7% normal male serum (hCG free) or both were also prepared. After a 15 minute incubation period, 50 pl of a dilute hCG-latex (Roche) suspension was added to each mixture.
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US597129 | 1984-04-05 | ||
US06/597,129 US4536478A (en) | 1984-04-05 | 1984-04-05 | Method for reducing non-specific interferences in agglutination immunoassays |
Publications (2)
Publication Number | Publication Date |
---|---|
EP0163393A2 true EP0163393A2 (fr) | 1985-12-04 |
EP0163393A3 EP0163393A3 (fr) | 1987-05-27 |
Family
ID=24390217
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP85302453A Withdrawn EP0163393A3 (fr) | 1984-04-05 | 1985-04-04 | Procédé de mise en oeuvre d'un essai immunologique d'agglutination |
Country Status (7)
Country | Link |
---|---|
US (1) | US4536478A (fr) |
EP (1) | EP0163393A3 (fr) |
JP (1) | JPS612078A (fr) |
CA (1) | CA1247523A (fr) |
DK (1) | DK149685A (fr) |
FI (1) | FI851326L (fr) |
NO (1) | NO851392L (fr) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0372413A2 (fr) * | 1988-12-02 | 1990-06-13 | BEHRINGWERKE Aktiengesellschaft | Milieu pour tests immunologiques comprenant un polymère contenant des groupes carboxyl |
FR2645967A1 (fr) * | 1989-04-12 | 1990-10-19 | Stago Diagnostica | Procede d'adsorption d'un materiel immunologique, utilisation dans le domaine des reactions antigene/anticorps |
WO2007111847A2 (fr) * | 2006-03-24 | 2007-10-04 | Aokin Ag | Utilisation d'additifs pour la réduction de liaison non spécifique dans des dosages |
Families Citing this family (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4695537A (en) * | 1982-05-21 | 1987-09-22 | The University Of Tennessee Research Corp. | Particles sensitized with detergent-treated antigen for agglutination immunoassay |
US4703001A (en) * | 1985-10-23 | 1987-10-27 | Synbiotics, Corporation | Immunoassay for the detection of serum analytes using pH dependent chastropic acids |
US4847209A (en) * | 1987-11-09 | 1989-07-11 | Miles Inc. | Latex agglutination immunoassay in the presence of hemoglobin |
US5100805A (en) * | 1989-01-26 | 1992-03-31 | Seradyn, Inc. | Quantitative immunoassay system and method for agglutination assays |
EP0535239A4 (en) * | 1991-03-18 | 1993-11-18 | Shiseido Company Limited | Collagen assaying method and kit |
US5484706A (en) * | 1993-05-19 | 1996-01-16 | Pasteur Sanofi Diagnostics | Immunoassay for analytes in samples using alkylating agents |
CA2127605A1 (fr) * | 1993-12-23 | 1995-06-24 | Peter J. Degen | Methode de separation par affinite |
CA2316130A1 (fr) * | 1999-08-19 | 2001-02-19 | Masanori Fukui | Methode de detection ou de determination de la presence d'antigenes capsidiques de l'hepatite c et reactif de detection ou de determination pour utilisation avec cette methode |
US7045098B2 (en) | 2001-02-02 | 2006-05-16 | James Matthew Stephens | Apparatus and method for removing interfering substances from a urine sample using a chemical oxidant |
US6890765B2 (en) * | 2001-11-02 | 2005-05-10 | Roche Diagnostics Operations, Inc. | Particles for immunoassays and methods for treating the same |
US6927071B2 (en) | 2001-12-07 | 2005-08-09 | Beckman Coulter, Inc. | Method for reducing non-specific aggregation of latex microparticles in the presence of serum or plasma |
US6777246B2 (en) | 2001-12-18 | 2004-08-17 | Roche Diagnostics Gmbh | Tertiary amine compounds for use in immunoassays |
US6984497B2 (en) * | 2002-04-05 | 2006-01-10 | Bio-Rad Laboratories, Inc. | Reducing non-specific binding in immunoassays performed on polymeric solid phases |
US20030022390A1 (en) * | 2002-05-30 | 2003-01-30 | Stephens James Matthew | Method and kit for making interfering substances in urine undetectable |
US7504200B2 (en) | 2007-02-02 | 2009-03-17 | Konica Minolta Medical & Graphic, Inc. | Photothermographic material |
JP5823377B2 (ja) * | 2010-03-31 | 2015-11-25 | 積水メディカル株式会社 | 測定系外成分による干渉を低減させる方法 |
JP5786188B2 (ja) * | 2010-06-08 | 2015-09-30 | 株式会社シノテスト | 試料中のc反応性蛋白質の測定試薬、測定方法及び測定範囲の拡大方法 |
US20160313310A1 (en) | 2013-12-13 | 2016-10-27 | Siemens Healthcare Diagnostics Inc. | Pretreatment agent in non-agglutination assays |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB2042170A (en) * | 1979-01-31 | 1980-09-17 | Takeda Chemical Industries Ltd | Reagent for tests utilizing latex agglutination |
EP0038181A2 (fr) * | 1980-04-15 | 1981-10-21 | TECHNICON INSTRUMENTS CORPORATION (a New York corporation) | Essais immunologiques par agglutination |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3689633A (en) * | 1969-01-13 | 1972-09-05 | Teikoku Hormone Mfg Co Ltd | Preparation of test sample for immunological assay of pregnancy of mares |
US3857931A (en) * | 1971-02-01 | 1974-12-31 | Hoffmann La Roche | Latex polymer reagents for diagnostic tests |
JPS5933228B2 (ja) * | 1978-12-25 | 1984-08-14 | 武田薬品工業株式会社 | 妊娠診断反応用被検液の前処理方法ならびに前処理剤 |
-
1984
- 1984-04-05 US US06/597,129 patent/US4536478A/en not_active Expired - Fee Related
-
1985
- 1985-04-02 FI FI851326A patent/FI851326L/fi not_active Application Discontinuation
- 1985-04-02 DK DK149685A patent/DK149685A/da not_active IP Right Cessation
- 1985-04-03 NO NO851392A patent/NO851392L/no unknown
- 1985-04-04 CA CA000478389A patent/CA1247523A/fr not_active Expired
- 1985-04-04 EP EP85302453A patent/EP0163393A3/fr not_active Withdrawn
- 1985-04-05 JP JP60072460A patent/JPS612078A/ja active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB2042170A (en) * | 1979-01-31 | 1980-09-17 | Takeda Chemical Industries Ltd | Reagent for tests utilizing latex agglutination |
EP0038181A2 (fr) * | 1980-04-15 | 1981-10-21 | TECHNICON INSTRUMENTS CORPORATION (a New York corporation) | Essais immunologiques par agglutination |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0372413A2 (fr) * | 1988-12-02 | 1990-06-13 | BEHRINGWERKE Aktiengesellschaft | Milieu pour tests immunologiques comprenant un polymère contenant des groupes carboxyl |
EP0372413A3 (fr) * | 1988-12-02 | 1991-07-03 | BEHRINGWERKE Aktiengesellschaft | Milieu pour tests immunologiques comprenant un polymère contenant des groupes carboxyl |
US5393659A (en) * | 1988-12-02 | 1995-02-28 | Behringwerke Aktiengesellschaft | Agent for immunochemical tests which contains polymers containing carboxyl groups |
FR2645967A1 (fr) * | 1989-04-12 | 1990-10-19 | Stago Diagnostica | Procede d'adsorption d'un materiel immunologique, utilisation dans le domaine des reactions antigene/anticorps |
WO2007111847A2 (fr) * | 2006-03-24 | 2007-10-04 | Aokin Ag | Utilisation d'additifs pour la réduction de liaison non spécifique dans des dosages |
WO2007111847A3 (fr) * | 2006-03-24 | 2007-11-22 | Aokin Ag | Utilisation d'additifs pour la réduction de liaison non spécifique dans des dosages |
US8124359B2 (en) | 2006-03-24 | 2012-02-28 | Aokin Ag | Use of additives for the reduction of non-specific binding in assays |
Also Published As
Publication number | Publication date |
---|---|
FI851326L (fi) | 1985-10-06 |
FI851326A0 (fi) | 1985-04-02 |
JPS612078A (ja) | 1986-01-08 |
DK149685A (da) | 1985-10-06 |
NO851392L (no) | 1985-10-07 |
CA1247523A (fr) | 1988-12-28 |
EP0163393A3 (fr) | 1987-05-27 |
US4536478A (en) | 1985-08-20 |
DK149685D0 (da) | 1985-04-02 |
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Legal Events
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17P | Request for examination filed |
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17Q | First examination report despatched |
Effective date: 19890202 |
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STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION HAS BEEN WITHDRAWN |
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18W | Application withdrawn |
Withdrawal date: 19900817 |
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R18W | Application withdrawn (corrected) |
Effective date: 19900817 |
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RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: RENO, JOHN M. Inventor name: SOKOLOFF, ROGER L. |